Background: For the determination of creatine kinase (CK)-MB, the immunoinhibition method is utilizedmost commonly. However, the estimated CK-MB activity may be influenced by the presence of CK isoenzymesin some conditions like cancer. Thus, a CK-MB-to-total-CK ratio more than 1.0 could be found in such asituation. The study aimed to explore the relationship of cancer to high CK-MB-to-total-CK ratio. Materialsand Methods: From January 2011 to December 2014, laboratory data on all CK-MB and total CK test requestswere extracted at Far Eastern Memorial Hospital (88,415 requests). Patients with a CK-MB-to-total-CK ratiomore than 1.0 were registered in this study. Clinical data including tumor location, tumor TNM stage andmetastatic status were also collected. Results: A total of 846 patients were identified with a CK-MB-to-total-CKratio more than 1.0. Of these, 339 (40.1%) were diagnosed with malignancies. The mean CK-MB-to-total-CKratio was significantly higher in malignancy than in non-malignancy (1.35±0.28 vs 1.25±0.23, p<0.001) groups.The most frequent malignancy with a CK-MB-to-total-CK ratio more than 1.0 was colorectal cancer (1.42±0.28,16.5%, n=56), followed by lung cancer (1.38±0.24, 15.9%, n=54) and hepatocellular carcinoma (14.5%, n=49).Higher CK-MB-to-total-CK ratios in hematological malignancies (1.44±0.41)were also noted. Additionally, theCK-MB-to-total-CK ratio was markedly higher in advanced stage malignancy than in early stage (1.37±0.26vs. 1.29±0.31, p=0.014) and significantly higher in liver metastasis than in non-liver metastasis (1.48±0.30 vs.1.30±0.21, p<0.001). Conclusions: The CK-MB-to-total-CK ratio is an easily available indicator and could beclinically utilized as a primary screening tool for cancer. Higher ratio of CK-MB-to-total-CK was specificallyassociated with certain malignancies, like colorectal cancer, lung cancer and hepatocellular carcinoma, as wellas some cancer-associated status factors such as advanced stage and liver metastasis.